pMarC9-R6k
(Plasmid #89477)

• Purpose: transposon plasmid, compatible with Tn-Seq
• Depositing Lab: George Church
• Publication: Lee et al Nat Microbiol. 2019 Apr 8. pii: 10.1038/s41564-019-0423-8. doi: 10.1038/s41564-019-0423-8.

Backbone

• Vector backbone: pBAM1
• Backbone manufacturer: Victor de Lorenzo lab
• Modifications to backbone: Tn5 replaced with marC9. Mariner transposon ends cloned into the transposon cassette. The transposon ends are mutagenized to introduce an mmeI cut site.

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Kanamycin, 100 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Pir1
• Growth instructions: Use BW29427 strain supplemented with 300uM DAP for this plasmid when conjugating from E. coli to Vibrio natriegens
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: pMarC9-R6k

Resource Information

• Supplemental Documents:
  • pMarC9_mmeI
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

BW29427 is an E. coli strain with diaminopimelic acid (DAP) auxotrophy. BW29427 growth requires 300uM DAP even when cultured in LB. Importantly, BW29427 does not grow in the absence of DAP, which simplifies counterselection of this host strain following biparental mating with Vibrio natriegens.

Please visit https://www.biorxiv.org/content/early/2016/06/12/058487 for bioRxiv preprint.